Erenna® IL-17A Immunoassay Kit (Cat# 03-0017-xx)

Introduction

The Erenna® IL-17A Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure IL-17A in plasma and serum samples. A capture antibody specific for IL-17A has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards, and samples into uncoated microplate wells. During incubation, the IL-17A present in the sample binds to the capture antibody on the coated MP. Unbound molecules are washed away during the subsequent buffer exchange and wash steps. Fluor-labeled detection antibody is added to each well and incubated. This detection antibody recognizes and binds to IL-17A that has been captured onto the MP. During the following wash step the MPs are transferred to a clean plate. Elution buffer is then added and incubated. The elution buffer dissociates the bound protein sandwich from the MP surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final microplate. The plate is loaded into the Erenna System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IL-17A present in the sample when captured. The amount of IL-17A in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-17A Immunoassay Kit¹
Lower Limit of Detection	0.01 pg/mL
Lower Limit of Quantification²	0.13 pg/mL
Upper Limit of Quantification	75 pg/mL
Low-end CV% Range	5 -7%
Low-end CV% Average	6%
Recommended Sample Volume	100 μL
Minimum Sample Volume Required³	50 μL

¹ See product insert for updated values
² LLoQ = 20% CV and ± 20% recovery
³ based upon median [IL-17A] in a healthy reference

FIGURE 1. [IL-17A] in EDTA plasma from 20 healthy donors, with median and interquartile range.

FIGURE 2. The Erenna® IL-17A Immunoassay kit low-end standard curve signal (left) and curve fit (right).

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Interleukin-17A (IL-17A) is the founding member of a group of cytokines called the IL-17 family. There are several members of the IL-17 family, from IL-17A to IL-17F (IL-17E is also called IL-25). IL-17A is involved in mediating proinflammatory responses by inducing the production of many other cytokines, chemokines, and prostaglandins from many cell types. The increased expression of chemokines attracts other cells, including neutrophils. IL-17A has many physiological functions, such as airway remodeling. As a result of these roles, the IL-17 family has been linked to many immune/autoimmune related diseases, including rheumatoid arthritis (RA), asthma, allergic responses, lupus, allograft rejection and antitumor immunity.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name:	Interleukin-17A
UniProtKB/Swiss-Prot ID:	Q16552
Protein-protein interaction database:	Q16552
Alternative Names:	Cytotoxic T-lymphocyte-associated antigen 8
Gene Names:	IL17A
Synonym Gene Names:	CTLA8, IL17
Function:	Induces stromal cells to produce proinflammatory and hematopoietic cytokines. Enhances the surface expression of ICAM1/intracellular adhesion molecule 1 in fibroblasts.
Biological Processes:	apoptosis, cell-cell signaling, immune response, inflammatory response, positive regulation of transcription from RNA polymerase II promoter, protein amino acid glycosylation
Molecular Function:	cytokine activity

* Source Info from www.uniprot.org.

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